Extracellular localization of the KpACE protein.

<p>Immunoblotting of culture supernatants and cell pellets was performed on <i>K</i>. <i>pneumoniae</i> cells carrying pkpACE (TH16356), which were cultured in an LB medium to OD<sub>600nm</sub> of 1.0. All samples were boiled for 10 minutes, and 15 μL of each sample was loaded onto a 12% SDS-PAGE gel. The KpACE-6×His fusion protein expressed by TH16356 was probed using a 6×His Tag monoclonal antibody (Invitrogen). The response regulator VicR, a known intracellular protein, was employed as a negative control to verify the localization of KpACE. Comparing the immunoblotting results of VicR and the KpACE-6×His fusion protein, we conclude that any detected signal for KpACE in the supernatant was not due to cell lysis or contamination with intracellular proteins.</p> <p>(PDF)</p>